More than 90% of advanced lung cancer patients will die within a year or two of diagnosis. Lung cancer will kill 160,000 Americans this year. Docetaxel is among the most effective chemotherapy drugs for this disease, but efficacy of chemotherapy alone for metastatic NSCLC has plateaued. Immunotherapy has produced tantalizing responses in a few NSCLC patients but has not been applicable widely. This proposal describes a novel approach to lung cancer immunotherapy that incorporates docetaxel chemotherapy to debulk the cancer, unmask hidden antigens and provide relative lymphodepletion with possible elimination of T regulatory cells combined with a novel autologous DRibble vaccine to stimulate an anticancer immune response. Our novel strategy targets T-cell responses to tumor-derived short-lived proteins by blocking proteasome-dependent protein degradation causing highly immunogenic tumor antigens to accumulate as ubiquitinated proteins in secretory vesicles of treated cancer cells. These vesicles, resembling apoptotic blebs, also contain short-lived proteins such as defective ribosomal products (DriPs) and are called DRibbles (DriPs in blebs). DRibbles can prime tumor-specific T cells in primary culture and induce strong tumor-specific T cells in vaccinated mice. Short-lived proteins are generally not cross-presented by host dendritic cells so peripheral tolerance to tumor- derived short-lived cancer proteins is less likely. However, short -lived proteins are highly expressed on tumor cells. This possibly explains why DRibble vaccines containing accumulated short-lived proteins were highly immunogenic and therapeutic in tumor-bearing mice. We hypothesize that a DRibble vaccine combined with lymphodepletion can overcome host tolerance to NSCLC cells and generate an immunologic and clinical anti-tumor response. The aims of our proposal are: 1) Perform a pilot clinical trial of autologous DRibble vaccine in patients with stage IIIB or IV NSCLC with malignant pleural effusion or subcutaneous metastases as a source of autologous cancer cells for vaccine generation. 2) Evaluate the immunologic efficacy of the vaccine by immune monitoring. Results from these studies will contribute to improved treatment for NSCLC by combining cytotoxic drugs and immunotherapy to maximize lung cancer control. Lung cancer is the most significant cancer killer in the Western world and is resistant to conventional chemotherapy with five-year survival in the 15% range. This proposal details a novel strategy employing an autologous tumor-derived vaccine (DRibble vaccine) plus chemotherapy that will exploit host and tumor factors to engender a powerful systemic anti-tumor immune response to tumor-associated antigens that are more likely to serve as tumor rejection antigens. This proposed strategy has the potential to improve outcome for thousands of lung cancer patients annually. PROJECT SUMMARY/ABSTRACT: More than 90% of advanced lung cancer patients will die within a year or two of diagnosis. Lung cancer will kill 160,000 Americans this year. Docetaxel is among the most effective chemotherapy drugs for this disease, but efficacy of chemotherapy alone for metastatic NSCLC has plateaued. Immunotherapy has produced tantalizing responses in a few NSCLC patients but has not been applicable widely. This proposal describes a novel approach to lung cancer immunotherapy that incorporates docetaxel chemotherapy to debulk the cancer, unmask hidden antigens and provide relative lymphodepletion with possible elimination of T regulatory cells combined with a novel autologous DRibble vaccine to stimulate an anticancer immune response. Our novel strategy targets T-cell responses to tumor-derived short-lived proteins by blocking proteasome-dependent protein degradation causing highly immunogenic tumor antigens to accumulate as ubiquitinated proteins in secretory vesicles of treated cancer cells. These vesicles, resembling apoptotic blebs, also contain short-lived proteins such as defective ribosomal products (DriPs) and are called DRibbles (DriPs in blebs). DRibbles can prime tumor-specific T cells in primary culture and induce strong tumor-specific T cells in vaccinated mice. Short-lived proteins are generally not cross-presented by host dendritic cells so peripheral tolerance to tumor- derived short-lived cancer proteins is less likely. However, short lived proteins are highly expressed on tumor cells. This possibly explains why DRibble vaccines containing accumulated short-lived proteins were highly immunogenic and therapeutic in tumor-bearing mice. We hypothesize that a DRibble vaccine combined with lymphodepletion can overcome host tolerance to NSCLC cells and generate an immunologic and clinical anti-tumor response. The aims of our proposal are: 1) Perform a pilot clinical trial of autologous DRibble vaccine in patients with stage IIIB or IV NSCLC with malignant pleural effusion or subcutaneous metastases as a source of autologous cancer cells for vaccine generation. 2) Evaluate the immunologic efficacy of the vaccine by immune monitoring. Results from these studies will contribute to improved treatment for NSCLC by combining cytotoxic drugs and immunotherapy to maximize lung cancer control. PROJECT NARRATIVE Lung cancer is the most significant cancer killer in the Western world and is resistant to conventional chemotherapy with five-year survival in the 15% range. This proposal details a novel strategy employing an autologous tumor-derived vaccine (DRibble vaccine) plus chemotherapy that will exploit host and tumor factors to engender a powerful systemic anti-tumor immune response to tumor-associated antigens that are more likely to serve as tumor rejection antigens. This proposed strategy has the potential to improve outcome for thousands of lung cancer patients annually.